pp1cb antibody  (Cell Signaling Technology Inc)

Journal: Antioxidants

Article Title: MnTE-2-PyP Suppresses Prostate Cancer Cell Growth via H 2 O 2 Production

doi: 10.3390/antiox9060490

Figure Lengend Snippet: Identification of MnTE-2-PyP-induced protein oxidation and PPPs activity measurements. ( A ) A representative image of Coomassie stained gels of non-oxidized proteins in PC3 cells. Proteins with reduced thiols were isolated by N-(biotinoyl)-N’-(iodoacetyl) ethylenediamine (BIAM) probe and selected regions of Coomassie gels were cut and sent for mass spectrometry analyses. ( B ) A representative image of a PP1CB western blot with ponceau staining in PC3 cells and corresponding densitometry analysis. ( C ) A representative image of a PP1CB western blot with ponceau staining in LNCaP cells and corresponding densitometry analysis. ( D ) A representative image of a PP1CB western blot with ponceau staining in PC3 and LNCaP cells and corresponding densitometry analysis. ( E ) PP1 activity measurement in PC3 cells with PBS or MnTE-2-PyP treatment. ( F ) Total PPP activity measurement in PC3 cells with PBS or MnTE-2-PyP treatment. ( G ) PP1 activity measurement in LNCaP cells with PBS or MnTE-2-PyP treatment. ( H ) Total PPP activity measurement in LNCaP cells with PBS or MnTE-2-PyP treatment. All data represent mean ± SD from at least three independent experiments. * p < 0.05 compared to PBS treatment.

Article Snippet: After blocking with 5% non-reduced fat milk in TBST for 1 h, the membranes were incubated overnight at 4 °C with the following primary antibodies: PP1CB (1:500), cyclin D1 (1:10,000), phospho-cyclin D1 (Thr 286, 1:1000), pRB, phospho-pRB (Ser780, 1:1000) (Cell Signaling Technology, Danvers, MA, USA) and p16 (1:5000), p21 (1:5000) (Abcam, Cambridge, MA, USA).

Techniques: Activity Assay, Staining, Isolation, Mass Spectrometry, Western Blot